Application of validamycin in preparation of chitinase inhibitor
A technology of Jinggangmycin and chitinase, applied in the fields of application, insecticide, biocide, etc.
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Embodiment 1
[0019] Example 1: Changes in regulation of trehalose metabolism after Spodoptera frugiperda injection of Validamycin
[0020] 1. Experimental materials: Spodoptera frugiperda comes from the breeding population of this laboratory, and the source of the tested insects comes from Zhejiang Academy of Agricultural Sciences. The feeding conditions of Spodoptera frugiperda are: temperature 26±1°C; photoperiod 16:8h (day: night), humidity 60±10%. The larvae of Spodoptera frugiperda were fed with artificial diet (Table 1), and the adults obtained nutrition with 10% honey water to simulate nectar.
[0021] Table 1 Feed formula for Spodoptera frugiperda larvae
[0022]
[0023]
[0024] 2. Microinjection of Spodoptera frugiperda Validamycin: Jinggangmycin injection with a concentration of 1 μg / μl was prepared with sterile water. Before injection, the volume of Jinggangmycin injection pumped out by each microinjector was first quantified with a standard capillary, and the volume o...
Embodiment 2
[0031] Example 2: The regulation of proteins related to chitin metabolism after Spodoptera frugiperda injection of Validamycin for 48 hours
[0032] 1. Changes of chitinase activity after Spodoptera frugiperda injection of Validamycin for 48 hours
[0033] Carry out chitinase activity assay according to the chitinase (Chitinase) kit specification sheet of Suzhou Branch Ming Biotechnology Co., Ltd., sample is the larva of Spodoptera frugiperda after microinjection 48h among the embodiment 1, assay method is as follows: Take about 0.1g of tissue, add 1ml of extract to homogenize in ice bath, then centrifuge at 10,000×g at 4°C for 20min, take the supernatant, put it on ice for testing, the measurement operation is shown in Table 2. Calculate the chitinase activity according to the absorbance value, the calculation formula is: chitinase activity (mg / h / g fresh weight) = (ΔA + 0.2753) ÷ 6.4108 × V reverse total ÷ (V sample ÷ V sample total × W )÷T×10=3.899×(ΔA+0.2753)÷W(V total: th...
Embodiment 3
[0040] Embodiment 3: Evaluation of the effect of Validamycin on the growth and development of insects
[0041] 1. Metamorphic development of Spodoptera frugiperda after injection of Validamycin
[0042] The Spodoptera frugiperda injected with 1 μg / μl Validamycin by the method of Example 1 was observed phenotype every 24 hours, and the abnormal phenotype was photographed with a camera (Canon, EOS 50D), and its pupation rate, eclosion rate, and mortality were counted every day . Figure 5 As shown, the pupation rate and eclosion rate of Spodoptera frugiperda injected with Validamycin had no significant difference compared with the control group, but the mortality rate injected with Validamycin began to be higher than that of the control group from the 4th instar and continued until the adult mortality were consistently higher than those in the control group. Compared with normal F. frugiperda, the number of F. frugiperda that died after injecting Validamycin was more, and the ...
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